Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris.
We analyzed and compared the physicochemical and immunochemical properties of recombinant human serum albumin (rHSA) from Pichia pastoris with those of plasma-derived human serum albumin (pHSA). The second virial coefficient of rHSA, obtained from colloid osmotic pressure measurements at pH 6.7 +/- 0.1 was not significantly different from that of pHSA (P > 0.05). A 25% rHSA solution exhibited Newtonian flow, and the viscosity of 25% rHSA at 20 +/- 0.02 degrees C was not significantly different from that of 25% pHSA (P > 0.05). We analyzed the long- and medium-chain fatty acid composition of rHSA by reverse-phase HPLC using 9-anthryldiazomethane as the fluorescent labeling reagent. The total amount of fatty acid was higher for pHSA than for rHSA. The fatty acid composition of the rHSA preparation was the same as that of the pHSA preparation. However, the amounts of palmitic acid (C16:0) and stearic acid (C18:0) in rHSA were much lower than those in pHSA. Interestingly, we found that P. pastoris produced linolenic acid (C18:3) because it was detected in rHSA. The immunochemical properties of rHSA were analyzed by a parallel line assay method using anti-pHSA polyclonal antibody, and were identical to those of pHSA (P > 0.05).
Ohtani
W
W
Research Division, Green Cross Corporation, Osaka, Japan.
Nawa
Y
Y
Takeshima
K
K
Kamuro
H
H
Kobayashi
K
K
Ohmura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000235:genetics; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003429:Cross Reactions; D005227:Fatty Acids / Q000032:analysis; D006801:Humans; D007120:Immunochemistry; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012449:Safety; D012709:Serum Albumin / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology
D000941:Antigens; D005227:Fatty Acids; D009842:Oligopeptides; D011994:Recombinant Proteins; D012709:Serum Albumin